Process of expanding T cells

Abstract

The present disclosure relates to a novel process for expanding T cells, such as autologous T cells, cell populations therefrom, pharmaceutical compositions comprising the said cell populations and use of the cells and compositions for treatment, particular the treatment or prophylaxis of virus infection and/or cancer, for example in immune compromised or immune competent human patients.

The invention claimed is: 
     
       1. A process for in vitro expansion of antigen specific T cells, comprising steps of:
 a) culturing a population of peripheral blood mononuclear cells (PMBCs) in the presence of:
 i) a peptide or a peptide mix relevant to one or more target antigens OR dendritic cells which have been pulsed with the peptide or the peptide mix, and 
 ii) at least one cytokine, thereby obtaining a population of T cells; and 
 
 b) culturing the population of T cells in the presence of:
 antigen presenting cells (APCs) which have been pulsed with the peptide or the peptide mix, wherein the APCs are selected from the group consisting of:
 (1) dendritic cells; and 
 (2) antigen presenting T cells autologous to the PMBCs; and 
 
 an artificial co-stimulatory factor, 
 
 
       characterized in that the process does not employ a live virus; a viral vector; or DNA or RNA encoding the target antigen(s). 
     
     
       2. The process according to  claim 1 , further comprising repeating step b) until a desired quantity of antigen-specific T cells is obtained. 
     
     
       3. The process according to  claim 1  wherein the population of PBMCs is cultured for 12 or fewer days. 
     
     
       4. The process according to  claim 1 , wherein the population of T cells is cultured for 12 or fewer days. 
     
     
       5. The process according to  claim 1 , which is performed in a vessel comprising a gas permeable culture surface. 
     
     
       6. The process according to  claim 1 , wherein the one or more target antigens are selected from the group consisting of antigens of an Epstein-Barr Virus, a Vaccinia Virus, and a Varicella Zoster Virus. 
     
     
       7. The process according to  claim 1 , wherein the peptide mix comprises between 2 and 1000 peptides. 
     
     
       8. The process according to  claim 1 , wherein the peptides are about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 amino acids in length. 
     
     
       9. The process according to  claim 1 , wherein the antigen is LMP1. 
     
     
       10. The process according to  claim 1 , wherein the antigen is LMP2. 
     
     
       11. The process according to  claim 1 , wherein the antigen is EBNAI. 
     
     
       12. The process according to  claim 1 , wherein the antigen is BARFI. 
     
     
       13. The process according to  claim 1 , wherein the at least one cytokine present in step a) is IL-4. 
     
     
       14. The process according to  claim 1 , wherein the at least one cytokine present in step a) is IL-7. 
     
     
       15. The process according to  claim 1 , wherein step b) further comprises at least one cytokine, wherein the at least one cytokine present in step b) is IL-15. 
     
     
       16. The process according to  claim 1 , wherein the artificial co-stimulatory factor is a cell engineered to express one or more co-stimulatory molecules on its surface. 
     
     
       17. The process according to  claim 16 , wherein the one or more co-stimulatory molecules are independently selected from CD80, CD86, CD83, OX-40 ligand and 41BB-ligand or a fragment thereof. 
     
     
       18. The process according to  claim 1 , wherein the cell comprises CD80, CD86, CD83, OX-40 ligand and 41BB-ligand. 
     
     
       19. The process according to  claim 1 , wherein the process is performed in a GRex™ system. 
     
     
       20. The process according to  claim 8 , wherein peptides in the peptide mix overlap by 2, 3, 45, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 or more amino acids. 
     
     
       21. The process according to  claim 1 , wherein the antigen specific T cells are allogeneic.